THE  BEHAVIOR  OF  SOME  AROMATIC 
ESTERS  OF  THERAPEUTIC  VALUE 
IN  METABOLISM 


BY 


RALPH  LESTER  HORST 

A.B.  Nebraska  Wesleyan  University,  1918 


THESIS 

SUBMITTED  IN  PARTIAL  FULFILLMENT  OF  THE  REQUIREMENTS 
FOR  THE  DEGREE  OF  MASTER  OF  SCIENCE  IN  CHEMISTRY 
IN  THE  GRADUATE  SCHOOL  OF  THE  UNIVERSITY 
OF  ILLINOIS,  1922 


URBANA,  ILLINOIS 


Digitized  by  the  Internet  Archive 

in  2015 


https://archive.org/details/behaviorofsomearOOhors 


UNIVERSITY  OF  ILLINOIS 


THE  GRADUATE  SCHOOL 


May  39 


1 HEREBY  RECOMMEND  THAT  THE  THESIS  PREPARED  UNDER  MY 

SUPERVISION  BY. Ralph  Lester  Korat 

entitled.  The  Behavior  of  Some  Aromatic  Esters  of 

Therapeutic  Value,  in  Metabolism. 

BE  ACCEPTED  AS  FULFILLING  THIS  PART  OF  THE  REQUIREMENTS  FOR 
THE  DEGREE  of  Master  of  Science 


In  Charge  of  Thesis 


Head  of  Department 


Recommendation  concurred  in* 


Committee 


on 


Final  Examination* 


Required  for  doctor’s  degree  hut  not  for  master’s 


■ 


■ 


.... 


Acknowledgement . 

The  writer  w ishes  to  take  this  opportunity 
to  express  his  appreciation  of  the  guidance  and 
help  given  by  Dr.  H.B.  Lewis  in  the  preparation 

of  this  thesis. 


fable  of  Contents 


page 

I.  Introduction 1 

II.  Historical. 1 

IIL  Experimental 4 

IV.  Discussion 7 

V Tables 10 

V.  General  Conclusion 9 

VI.  Summary 9 

VLL  Bibliography • 22 


. 


1 


I. 

Introduction 

In  the  previous  studies  of  benzyl  alcohol  and  its  esters,  tv/o 
chief  purposes  have  been  in  mind.  I.  The  uses  and  action  of  benzyl 
alcohol  as  a local  anesthetic.  II.  The  physiological  action  of  the 
benzyl  esters. 

Since  the  use  of  this  alcohol  and  its  esters  is  increasing  in 
therapeutical  practices,  it  was  considered  of  importance  to  study 
the  behavior  of  these  compounds  in  metabolism  in  the  animal  body. 

II. 

Historical 

In  1914,  Filippi,  (1),  an  Italian  investigator,  found  that 
benzyl  alcohol  caused  sleepiness,  a paralysis,  and  a lowering  of 
the  body  temperature. 

Ilacht  and  his  co-workers  have  made  many  studies  on  this  alco- 
hol and  its  esters  and  it  is  to  him  that  we  owe  most  of  our  know- 
ledge of  their  action. 

His  first  work  was  carried  out  in  1918  (2)  with  the  alcohol. 

He  found  that  it  is  a powerful  local  anesthetic  and  used  it  with 
great  success  in  the  treatment  of  abscesses,  extraction  of  teeth 
and  skin  grafting.  A 1 cJo  aqueous  solution  was  used.  He  also  used 
it /as  a local  anesthetic  in  subcutaneous  and  inti®,  muscular  injec- 
tions. He  found  that  it  had  an  irritating  effect  upon  the  skin, 
but  that  it  was  an  antiseptic  and  the  wound  healed  well.  Eucaine 
cocaine  and  novocaine  did  not  have  antiseptic  properties.  (4) 

Ilacht,  Satani , and  Schwartz  (5)  even  stated  that  benzyl  alcohol  was 
antiseptic  and  germicidal  to  gonococcus,  staphylococcus  and  B.  6oli. 

Ilacht  also  found  that  it  ?/as  very  useful  as  a local  anesthetic 


> 

. : 


. '• 


2. 


in  genito-ur inary  work.  (6)  Sollman  (7)  studied  its  anesthetic  ac- 
tion upon  various  types  of  mucous  membrane  and  found  it  to  he  weak- 
er in  its  action  than  cocaine. 

Voegtlin  and  Livingstone  (8)  injected  it  in  regions  near  the 
spinal  cord  and  in  this  way  anesthetized  the  posterior  part  of  the 
body.  They  also  obtained  a lowering  of  blood  pressure  and  a depres 
sion  of  respiration. 

Macht  (6)  found  that  benzyl  benzoate  and  benzyl  acetate  (2) 
both  had  the  same  ant i spasmodic  or  anesthetic  effect  upon  muscles 
as  papaverine,  and  had  a lower  toxicity.  He  stated  that  they  were 
easily  metabolized  and  considers  the  action  of  papaverine  to  be  due 
to  the  benzyl  group  present.  Both  could  be  used  in  excessive  per- 
istalsis. He  found  the  acetate  bo  be  of  the  higher  toxicity  due 
to  its  bein^  absorbed  more  rapidly.  The  benzoate  was  found  to  be 
useful  in  spasmodic  dysmenorrhea,  spastic  conditions  of  the  intes- 
tine and  stomach,  threatened  abortion  and  renal  and  biliary  colic. 
It  also  had  a depressing  action  upon  blood  pressure.  Mason  (9) 
also  found  this. 

Heller  and  Steinfield  (10)  found  that  the  injected  benzoate 
had  no  toxic  action  upon  th6  leucocytes. 

Ilielson  and  Higgins  ( n)  injected  the  cinnamate  and  benzoate. 
Both  caused  a relaxation  of  the  intestine  and  a lowering  of  blood 
pressure,  also  a venous  coloration  of  the  arterial  blood.  Later 
(12)  they  used  other  benzyl  esters  and  they  all  produced  the  same 
general  effect.  -They  considered  the  action  to  be  due  to  the  hydrol 
ysis  Of  the  ester  to  benzyl  alcohol.  In  simple  esters  the  action 
was  proportional  to  the  rate  of  hydrolysis.  In  substituted  esters 
as  the  salicylate  and  acetyl-salicylate  the  action  was  more  rapid. 


3. 

Macht  (13)  also  studied  the  action  of  benzyl  succinate  and 
found  it  to  have  the  sarae  toxicity  as  the  benzoate,  but  the  action 
as  an  ant spasmodic  was  milder  and  longer  in  duration  due  to  the  slow- 
er breakdown  of  the  ester. 

Volwiler  and  Vliet  (14)  worked  with  various  benzyl  esters  and 
stated  that  the  antispasmodic  action  was  due  to  the  nucleus  ring. 
They  found  the  rate  of  hydrolysis  with  ZOH  to  increase  in  the  fol- 
lowing order : salicylate,  benzoate,  stearate,  cinnamate,  acetate, 
succinate  and  fumarate. 

Shonle  and  Row  (15)  prepared  a variety  of  benzyl  esters  of  the 
saturated  and  unsaturated  higher  fatty  acids.  'They  found  them  to 
be  insoluble  in  water,  but  readily  soluble  in  chloroform,  ether  and 
benzene.  Because  of  their  low  solubility  they  had  neither  local 
anesthetic  effect  nor  irritant  action  upon  mucous  membrane.  Clin- 
ical tests  demonstrated  their  antispasmodic  action  in  asthma,  dys- 
menorrhea, high  blood  pressure,  pylorospasm  and  spastic  constipation. 

Although  not  hydrolyzed  by  steam  these  esters  were  readily  split 
by  lipase.  If  aromatic  acids  were  substituted  for  the  aliphatic, 
the  hydrolysis  was  much  slower.  The  laurate,  myristate.,  palmitate , 
stearate,  oleate,  lactate  and  the  m-  and  p-  arr,ino  benzoates  were 
prepared.  Of  these  only  the  m-  and  p-  ar ino  benzoates  had  local 
anesthetic  properties. 

The  work  of  these  investigators  shows  the  therapeutic  value  of 
benzyl  alcohol  and  its  esters  as  local  antiseptics  and  as  antispas- 
modics . 

The  work  on  the  metabolism  of  benzyl  alcohol  and  its  esters  is 
very  limited. 

Macht  (2)  investigated  the  fate  of  the  acetate  and  the  benzoate 


4 


in  the  body  and  found  that  they  were  in  a large  measure  excreted  as 
hippuric  acid  in  the  urine.  This  transformation  of  these  esters 
into  hippuric  acid  he  considered  as  involving  a transition  from 
the  esters  through  the  alcohol.  In  rabbits  this  transformation  was 
more  rapid  than  in  the  dog.  His  work  on  the  alcohol  would  also  in- 
dicate that  it  is  in  a large  measure  excreted  in  the  urine  in  the 
form  of  hippuric  acid. 

III. 

Experimental . 

This  work  was  carried  out  in  order  to  study  the  behavior  of 
benzyl  succinate  in  metabolism,  as  compared  with  other  benzyl  enters 
and  30dium  benzoate,  making  use  of  the  increased  elimination  of 
hippuric  and  free  benzoic  acid  in  the  urine  as  a guide  to  the  met- 
abolism of  the  compound. 

The  purity  of  the  succinate  was  determined  by  saponification 
with  a known  amount  of  KOH.  It  was  found  to  be  99.4-100$  benzyl 
succinate . 

It  was  first  decided  to  study  the  action  of  hog  liver  lipase 
upon  this  ester,  a di-carboxylic  ester,  as  compare!  with  a mono-car- 
boxylic ester,  ethyl  propionate,  by  the  method  which  Christman  and 
Lewis  (16)  used  in  their  lipase  studies.  A glycerol  extract  of 
hog  liver  was  made  within  one  hour  after  the  death  of  the  animal. 
Since  benzyl  succinate  is  insoluble  in  water  and  does  not  melt  at 
body  temperature,  emulsions  with  dextrin  and  acacia  were  tried  with 
continuous  shaking  in  a machine,  but  no  splitting  of  the  ester  by 
the  lipase,  worthy  of  mention  could  be  obtained.  This  seemed  to  be 
due  to  a poor  emulsion  as  the  enzyme  was  found  to  be  very  active. 

Attention  was  then  turned  to  the  administration  of  the  ester 


■ 


. 


■ ’ 


5. 


per  os  and  studying  its  splitting  in  the  animal  body.  The  ester 
might  have  been  split  to  the  alcohol  which  could  have  been  elimina- 
ted in  the  urine  as  such  or  oxidized  and  eliminated  as  hippuric 
acid,  or  passed  through  into  the  urine  unchanged. 

It  was  decided  to  use  the  method  of  Kingsbury  and  Swanson  (16) 
for  the  determination  of  hippuric  acid  in  the  urine  and  thereby 
estimate  the  amount  of  ester,  which  was  split,  oxidized  and  elim- 
inated as  free  benzoic  or  as  hippuric  acid.  This  method  is  really 
a determination  of  total  benzoic  acid  or  free  benzoic  plus  hippuric 
acid.  From  the  work  of  others  this  was  known  to  be  a very  accurate 
method.  It  consists  of  the  hydrolysis  of  the  hippuric  acid  with 
sodium  hydroxide,  oxidation  of  interfering  substances  with  nitric 
acid  and  potassium  permanganate^ extract ion  of  the  benzoic  acid  with 
chloroform,  and  titration  of  this  extract  with  standard  sodium  eth- 
ylate . 

There  is  a possibility  her6  that  any  free  benzyl  ester  in  the 
urine  would  be  hydrolyzed  and  the  alcohol  oxidized  to  benzoic  acid 
by  the  nitric  acid  and  potassium:  permanganate  and  titrated  as  ben- 
zoic acid  present  in  the  urine  originally.  In  order  to  determine 
whether  this  happens, this  method  was  used  upon  a known  amount  of 
the  ester.  A negligible  amount  was  changed  to  benzoic  acid  in  the 
procedure.  (Table  I). 

If  part  of  the  ester  is  hydrolyzed  to  alcohol  and  eliminated 
as  such  in  the  urine,  there  is  a possibility  of  this  being  oxidized, 
in  this  procedure,  to  benzoic  acid  and  determined  as  benzoic  acid 
originally  present.  This  method  was  also  tested  on  urine  contain- 
ing a known  amount  of  the  alcohol.  ITo  appreciable  oxidation  of  the 
alcohol  was  obtained  in  this  procedure.  (Table  Ilj. 


.»U 


• • 

• • 

■ 


« 


5 • 

For  the  determination  of  free  benzoic  acid  or  benzoates  in  the 


urine,  the  method  of  P.aiziss  and  Dubin  (17)  was  used.  This  invol- 
ves acidification  with  a small  amount  of  nitric  acid  and  extraction 
of  the  benzoic  acid  with  toluene.  Since  benzyl  succinate  is  also 
soluble  in  toluene,  it  was  decided  to  determine  its  presence  in  the 
urine  by  saponification  of  the  neutralized  toluene  extract  with  ex- 
cess sodium  ethylate  and  titration  of  the  excess. 

The  complete  procedure  to  be  used  was  carried  out  upon  a urine 
containing  a known  amount  of  added  sodium  hippurate,  sodium  benzo- 
ate and  benzyl  succinate  dissolved  in  alcohol.  An  aliquot  was  used 
for  the  determination  of  total  benzoic  acid  or  hippuric  acid  plus 
free  benzoic  acid  from  the  benzoate  added.  A good  recovery  of  these 
two  compounds  was  obtained. 

Another  aliquot  was  tested  for  free  benzoic  acid  and  after  the 
neutralization  of  the  toluene  extract,  an  excess  of  sodium  ethylate 
was  add6d  and  this  refluxed  to  saponify  the  ester.  The  remaining 
alkali  was  titrated  and  the  amount  of  saponifiable  matter  determined 
A good  recovery  of  the  sodium  benzoate  and  benzyl  succinate  was  ob- 
tained. (Table  III). 

In  the  following  experiments  rabbits  fed  a standard  diet  were 
used  as  the  experimental  animals.  A weighed  amount  of  the  alcohol, 
ester  or  sodium  benzoate  was  administered  per  os,  the  bladder  emp- 
tied at  various  intervals  and  the  urine  analyzed. 

Benzyl  succinate  melts  at  45* C.  and  was  given  melted  and  emul- 
sified with  milk  as  f -as  possible.  Benzyl  alcohol  and  the  ace- 
tate, being  liquids,  were  well  mixed  with  milk  and  given  in  that 
form.  Sodium  benzoate  was  dissolved  in  water  and  given,  mixed  with 
a small  amount  of  milk. 

The  normal  diet  used  was  10  grams  of  sucrose  in  150  cubic  cen- 


7 


timeters  of  milk  in  the  first  expert)  ents  with  10-15  grains  of  oats 

in  addition  in  the  later  work, 
was 

flie  urin6  collected  as  indicated.  Three  days  on  a normal  diet 
elapsed  between  the  feeding  of  the  compounds,  but  this  was  varied 
later  as  shown  in  the  tables.  The  number  of  cubic-centimeters  of 
standard  sodium  ethylate  for  saponification  of  the  neutralized  tol- 
uene extract  was  also  recorded.  Combined  benzoic  acid  is  calculated 
from  the  difference  between  total  and  fre6  benzoic  acid.  In  order 
to  make  a direct  comparison  of  these  possible,  equivalent  amounts 
were  us6d. 

The  urine  in  each  case  was  diluted  to  a definite  volur.i6  and 
an  aliquot  used  for  each  determination.  Benzyl  succinate  was  made 
the  standard  in  the  amounts  of  the  compounds  used.  1£;1.5,  and  2.0 
grams  of  this  ester  were  used  according  to  the  comparative  weight 
of  the  animal,  and  equivalent  amounts  of  the  other  compounds  cal- 
culated from  these  values. 

IV. 

Discussion 

As  shown  by  these  tables,  the  elimination  of  these  compounds 
fed,  varies  greatly  with  the  individual  animal.  As  we  are  most  in- 
terested in  the  comparison  of  the  succinate  with  the  other  compounds 
its  relation  is  the  most  important. 

In  table  IV  and  V we  find  it  elii  inated  more  slowly  than  the 
other  compounds  but  its  recovery  was  more  complete.  In  table  V,  it 
was  eliminated  at  the  same  rate  as  sodium  benzoate,  but  its  recovery 
was  much  more  complete.  Here  it  is  also  eliminated  more  rai^idly 
than  the  alcohol  or  acetate.  Yet  in  tables  VII  and  Ik  , the  elim- 
ination was  very  3low,  and  not  as  complete  a recovery  as  of  the  al- 


Q. 

cohol  or  acetate  was  obtained,  although  the  recovery  of  these  was 
very  incomplete. 

In  tables  VIII  and  IX  with  feeding  the  succinate  only,  we  find 
the  elimination  greater  during  the  first  eight  hours  in  one  case  and 
greater  during  the  last  sixteen  hours  in  the  other.  The  greater 
elimination  of  the  succinate  in  the  first  eight  hours  is  also  found 
in  table  XI. 

In  table  IV,  X and  XII,  sodium  benzoate  was  eli]  inated  most  ra- 
pidly, but  in  table  XII  more  of  the  succinate  was  recovered.  Ben- 
zyl acetate  was  the  most  completely  recovered  of  these  compounds  in 
tabl6  X,  while  it  was  the  slowest  to  be  eliminated  in  tabl6  XII. 

In  the  latter  table  there  was  also  a slow  elimination  of  the  succi- 
nate but  faster  than  that  of  the  alcohol  and  more  complete  than  the 
acetate  or  sodium  benzoate. 

The  low  results  in  tables  XI  and  XII  may  be  due  to  a poor  nu- 
tritive condition  of  the  animal,  later  resulting  in  death. 

In  tables  IV,  V and  VI,  are  given  the  amounts  of  alkali  requir- 
ed for  saponification  of  the  sample.  This  determination  was  omitted 
in  the  subsequent  experiments,  since  it  was  practically  constant  af- 
ter feeding  these  compounds. 

In  tables  XI  and  XII  the  free  benzoic  acid  determination  was 
omitted  since  it  also  remained  almost  constant  during  the  feeding 
of  these  derivatives.  The  amount  of  benzoic  acid  eliminated,  wheth- 


er free  or  combined  is  of  interest  in  this  work,  mor6  than  the  form 
of  the  benzoic  acid. 


■ 

. 


, 


' 


V 


9 


V. 

General  Conclusion 

Benzyl  succinate  is  alowly  But  completely  eliminated  in  the 
urine  as  hippuric  or  fre6  benzoic  acid. 

VI. 

Summary 

I.  An  attempt  was  made  to  study  the  action  of  hog  liver  lipase 
upon  benzyl  succinate,  but  due  to  the  insolubility  of  the  ester,  and 
the  failure  to  obtain  a good  emulsion,  no  definite  results  were  ob- 
tained. 

II.  The  reagents  used  in  the  determination  of  total  benzoic  acid 
when  tested  on  benzyl  succinate  or  on  benzyl  alcohol  caused  only 
slight  oxidation  to  benzoic  acid. 

III.  Benzyl  alcohol,  acetate  and  succinate  and  sodium  benzoate 
wer6  fed  to  rabbits  and  the  rate  and  completeness  of  the  elimination 
of  the  hippuric  acid  in  the  urine  was  studied  and  compared. 

IV.  The  tables  indicate  that  benzyl  succinate  is  hydrolyzed  in 
the  animal  body  more  slowly  than  the  liquid  or  soluble  esters,  yet 
the  recovery  as  benzoic  acid  seems  to  be  the  most  complete. 

V.  There  was  no  increase  of  saponifiable  matter  in  the  urine 
after  feeding  this  ester.  This  indicates  that  that  ester  is  not 
absorbed  or  retained  as  such,  but  in  the  form  of  its  cleavage  pro- 


ducts 


Effect  of 

Table  I. 

the  procedure  for  hippuric  acid 

10. 

upon  benzyl  succinate. 

Benzyl  succi- 
nate taken. 

Benzyl  succi- 
nate equiva- 
lent to  ben- 
zoic acid. 

Benzoic  a- 
cid  found. 

3enzyl  succi- 
nate oxidized 
to  benzoic  a- 
cid . 

g* 

mg. 

mg. 

55 

2.0000 

1640.0 

1074.0 

6.5 

1.0000 

820.0 

52.00 

6.3 

.6470 

530.0 

30.00 

5.7 

.5000 

410.0 

31.70 

7.7 

.2419 

200.0 

18.00 

9.0 

.2000 

164.0 

9.44 

5.7 

.1915 

157.0 

18.30 

11.7 

.1511 

123.7 

14.60 

11.8 

Table  II. 

Effect  of 

the  procedure  for 

hippuric  acid 

upon  benzyl  alcohol. 

Benzyl  alco- 

Benzyl  alco- 

Benzoic  Extra 

ben-  Benzoic  alee- 

hoi  added  to 

hoi  equiva- 

acid  zoic 

acid  hbiaoxidized 

urin6 . 

lent  to  ben- 

found  found 

to  benzoic 

zoic  acid. 

acid . 

8 • 

mg. 

mg.  mg. 

$ 

0> 

0 

36  • 6 . 

0 

0 

36.6  

— 

.800 

903.7 

54.0  17.4 

1.9 

.800 

903.7 

48.2  11.6 

1.3 

11 


Table  III. 

Recovery  of  compounds  added  to  urine  by  methods  used  upon 


experimental  urine. 


Experiment . 

BE1TZ0IG  ACID 

Added  as 

i 

' Recovered. 

I 

f 


I. 

mg. 

Sodium  benzoate  84.7 
Benzyl  succinate  81.9 
Hippuric  acid  34.1 

mg. 

88.0 

97.8 

70.3 

Jo 

103.9* 

119.5* 

206.0* 

Sodium  benzoate  100.0 

96.0 

96.0 

II. 

Benzyl  succinate  82.0 

76.8 

93.7 

Hippuric  acid  34.1 

66.7 

195.5* 

Hippuric  and  benzoic  acids  and  saponifiable  matter  are 
known  to  be  present  in  the  urine  and  corrections  were  not  made  for 
these  amounts,  hence  the  percentages. 


V - 


* 


* ' F 


:hu. 


12. 

Table 

IV. 

Rabbit  I. 

--male 

weight  = 2.46  kg 

Daily 

diets  150  go.  milk  and 

10  g.  sucrose 

and  10  g.  oats. 

Per- 

Dura- 

Benzyl 

Benzoic  Acid  I;  Alkali 

Benzyl  der- 

Benzyl 

iod. 

tion 

Der iva 

- To-  F 

ree  Com  for  sap- 

ivative  fed. 

compound 

tive 

tal 

bined.  onifica- 

calculated 

recover- 

used 

tion  of 

as  behzoic 

ed  as  ben 

toluene 
extract . 

acid. 

zoic  acid 

I. 

hrs . 
72 

(24)* 

mg. 

IPS* 

139.6 

(46.5) 

mg. 

16.0 

(5.2) 

ms.  cc. 

123.6  12.8 

(41.3)  (4.3) 

mg. 

/ 0 

II. 

24 

Benzyl 

succi- 

nate 

1000 

634.4 

31.6 

602.8  8.8 

819 

71.8 

III. 

24 

— 

205.0 

2.1 

202.9  3.2 

19.4 

IV. 

72 

__ 

224. 

5.1 

218.9  6.2 

* • 

(24)* 

- 

( 75) 

( 1 . 7 ) ( 72 . 6 ) (2.1) 

_ 

V. 

24 

Benzyl 

alcohol 

720.1 

5.1 

714.9  4.5 

833.4 

80.8 

VI. 

24 

(737.8) 

86.4 

3.4 

83.0  4.1 

4.8 

VII. 

24 

~ 

66 .4 

3.4 

63.4  4.9 

2.4 

VIII. 

48 

.. ' ' e 

108.6 

5.7^102.9  6.96 

(24)* 

(54.3) 

(2.9) 

(51.4)  (3.  5) 

IX. 

24 

2SS5&,**.  a. 

3.4 

705.4  3.3 

823.6 

80.4 

1012.6 

X. 

24 

— 

87.2 

3.4 

83.8  3.0 

_ 

4.94 

XI. 

24 

. — _ 

60.8 

3.4 

57.4  3.8 

__ 

4.6 

XII. 

48 

(122 

3.4 

118.6  3.4 

(24)* 

( 61) 

(1.7) 

(59.3)  (1.7) 

— 

— 

* Average  per  24  hours. 

13. 

Table  V. 

Rabbit 

2. 

male 

weight  2.60 

kg. 

Daily  diet 

: 150  cc. 

milk  and  10  g.  sucrose  and  10 

g.  oats. 

Benzyl 

BENZOIC  ACID 

alaka- 
10  li  for 

Benzyl 

deriva- 

Benayl 

canpoun* 

Per- 

iod 

Dura- 

tion 

deriva- 

tive 

used 

To-  Free  Combined 
tal 

saponifi- 
cation of 
toluene 
extract 

tive  fed 
calcula- 
ted as 
benzoic  acid 

recovered 
as  ben  - 
zoic 
acid 

hrs. 

mg. 

mg . mg • mg . 

cc . 

mg. 

P 

I 

72 

__ 

218.6  5.1  215.5 

11.08 

_ 

_ 

(24) 

* 

( 72.9  )( 1.7  )(  71.2) 

(3.7) 

II. 

24 

Benzyl 

acetate 

1109.0 

751.2  7.2  744.0 

5.52 

902 

75.2  ; 

III. 

24 

156.8  16.8  120.0 

4.8 

7.0 

IY. 

24 

l 

61.0  15.6  47.4 

4.7 

V. 

48 

101.7  17.8  85.9 

6.28 



(24) 

* 

( 50.9)  (8.9)  (42.0) 

3.14 

VI. 

24 

Benzyl 

succi- 

nate 

1000 

664.  5.1  658.9 

4.19 

819 

74.9 

VII. 

24 

152.7  5.1  147.6 

5.0 

12.4 

VIII. 

24 

— 

71.2  5.4  67.8 

4.11 

— 

2.5 

* Average  per  24  hours. 

14. 

Table 

VI. 

Rabbit 

3 male 

weight  2.54  kg. 

Daily 

diet:  15Q  cc. 

milk  and  10  g 

. sucrose 

and  15  g. 

oats . 

Ben  zyl 

B3UZ0IC 

ACID 

[ alaka- 
10  li  for 
aaponifi- 

Benz  yL 
da?iva- 
tive  fed 

Benzyl 
c ompound 
recovered 

Per- 

iod 

deriva  - 
Dura-tive 
tion  used 

m 

Tbtal  Free 

Combined 

cation  of 

toluene 

extract 

oakuliar- 
tod  as 
bsnz>ic  a 

as  benzo- 
ic acid 

hrs  mg. 

mg. 

mg . 

mg. 

cc . 

mg. 

/U 

I. 

72  

183.2 

2uQ 

163.2 

10.88 

_ 

__ 

(24)*  _ 

(613 

(6.7) 

(54.4) 

(3.63) 

. r_ 

II. 

24  Bersyl 
succi- 
nate 
2000 

1375.2 

39. 

1337.2, 

4.51 

1639 

80.3 

III. 

24  

132.8 

10.1 

122.7 

2.64 

4.38 

IV. 

24  

122.2 

3.9 

118.3 

3.87 

3.72 

V. 

72  

152.6 

67.8 

84.  % 

5.96 

(24)*  _ 

(50P) 

(22 .6) 

(28.3 

(1.99) 

VI. 

24  Benzyl 

acetal  e 
2013.5 

1236. 

20.8 

1215.2 

2.24 

1637. 

7 72.3 

VII. 

24  __ 

135.2 

20.8 

114.4 

2.40 

_ 

5 J.  5 

VIII. 

24  

'71.  5 

6.9 

64.6 

2.24 

__  . 

1.60 

IX. 

48  

109.0 

15.6 

93.4 

6.0 

(24)* 

(54.3 

(7.8) 

(46.7) 

(3.0) 

X. 

24  Benz  yl 

alcchoL 
1828.0 

1512. 8 

2.8 

15  ia 

2.56 

2065 

70.5 

XI. 

24  

134.7 

5.2 

129.5 

2.0 

3.88 

XII. 

24  

78.1 

3.5 

74.6 

2.8 

_ 

1.14 

XIII. 

48  

92.8 

7.8 

85.0 

5.36 

— 

— 

Table 

VI.  (cont 

. ) 

(24)*  _ 

(46.4) 

(3.9) 

(42.5) 

(1.68) 

, - 

- 

XIV. 

24  Sodium 
benzoate 
1933 

1376. 

8.8 

1367.2 

3.04 

1637.1 

81.2 

XV. 

24  

62.5 

5.2 

57.3 

2.67 

_ 

.99 

XVI. 

24  

73.2 

5.2 

68.0 

2.40 

LS4 

XVII. 

48  

93.6 

13.2 

8014 

3.8 

(24)*  __ 

(46.8) 

(6.6) 

(40.2) 

(1.9) 

— 

— 

* Average  p6r  24  hours. 


16 


Table  VII. 


Rabit 

4 

male 

weight:  2.06 

kg. 

Daily 

diet : 

150  cc 

. milk 

and  10  g.  sucrose  and  15  g.  oats. 

Per- 

iod 

Dura- 

tion 

Wt. 

Benzyl 

deriv- 

ative 

used 

Benzoic 
To-  Dree 
tal 

Acid 

Com- 

bined 

B al.  Ben. 
10  der. 

for  sap*  fed 
onifi.  cal. 
of  tol.  ben- 
extract  zoic 

Benzyl 
c omp . 
recov- 
ered as 
benzoic 
acid. 

hrs . 

kg. 

mg. 

mg.  mg. 

mg. 

c c . mg . 

fo 

I 

96 

2.04 

-- 

536.8  238.5 

298.3 

5.95 

-- 

( £4)* 

-- 

-- 

( 134. 2)(.  59.6) 

(74.6) 

(1.49) 

-- 

II. 

£4 

2.08 

Benzjfc. 
sue . 
1500 

525.6  1.7 

323.9 

2.11  1228.5 

15.6 

III. 

24 

2.08 

-- 

685.6  5.2 

680.4 

2.67 

45.0 

IV. 

24 

^ • _L 

— 

122.  5.2 

116.8 

2.67 

--- 

V. 

48 

2.10 

-- 

301.6  10.4 

291.2 

4.4 

-- 

(24)* 

— 

— 

(150.8)  ( 5.2  ) ( 145.6) 

(2.2) 

-- 

VI. 

24 

2.10 

Benzyl 

acetat 

1511.7 

980.9  17.6 

e 

963.3 

2.24  1219.5 

68.2 

VII. 

24 

2.13 

-- 

73.6  7.2 

66.4 

2.24 

-- 

VIII. 

24 

2.13 

-- 

98.6  5.2 

93.4 

2.4 

-- 

IX. 

48 

2.18 

-- 

192.3  10.4 

181.9 

3.36 

-- 

(24)* 

— 

-- 

(96.2)  (5.2) 

(91.0) 

(1.68) 

-- 

X. 

24 

2.17 

Benzyl  1005.3  10.4 
alcohol. 

1087.3 

994.9 

2.1  1 228.3 

74.1 

XI. 

24 

2.16 

-- 

93.7  5.2 

88.5 

2.4 

-- 

XII. 

24 

2.19 

-- 

73.2  5.2 

68.0 

2.11 

-- 

XIII  . 

48 

2.25 

-- 

145.6  21.2 

124.4 

3.16 

-- 

(24)* 

-- 

— 

(72.8)  (10.6) 

(62.3 

(1.58) 

-- 

* 


Average  per  24  hours 


, 


. 


. 


«■ 


- 


•>  ,j 

* 


4 


' 


t 


1 


17. 

Cable  VIII 

Rabbit 

6 male 

weight : 

2.12  kg. 

! Daily 

diet : 

150  cc.  milk 

and  10  s:.  sucrose  and  15  sr  oats 

Per- 

iod 

Dura- 

tion 

- Benzyl  de-  Benzoic  Acid 

rivative  used  To-  Tree  Com- 
tal  bined 

Benzyl  de- 
rivative 
used,  cal- 
culated as 
benzoic  acii 

Benzyl  de 
rivative 
recovered 
as  benzoic 
acid. 

hrs 

mg. 

mg.  mg.  mg. 

mg. 

$ 

I. 

24 

85.7  15.9  69.7 

— 

-- 

II. 

8 

benzyl  sue 

cinate 

1500 

- 315.0  — 

1228.2 

18.68 

III.  * 

16 

732.0  -- 

52.80 

Rabbit 

7 

male 

Cable  12 

weight:  2 

.6  kg. 

\ Daily 

diet : 

150  cc.  milk 

and  10  g.  sucrose  and 

-L  • • 0 ctlj  S • 

I. 

24 

-- 

124.2  — 

-- 

. 

II. 

8 

benzyl  suc- 
cinate 

2000 

995.4  23.4  972.0 

1637.6 

53.5 

III. 

16 

-- 

336.7  3.9  332.8 

— 

13.0 

IV. 

24 

-- 

212.3  3.9  208.4 

— 

5.38 

V. 

* 

24 

— 

94.2  3.9  90.3 

— 

— 

* Died 

18. 

m 

able  X. 

Rabbit  8 

male 

weight:  2 

.06  kg. 

Daily 

diet 

: 150  cc. 

milk  and 

10  g. 

sucrose 

and  15  g.  oats 

Per- 

iod 

Dura- 

tion 

Benzyl  de- 
rivative 
used 

Benzoic 
Total  Free 

Acid 

Combined 

Benzyl  deri- 
vative used 
calculated 
as  benzoic 

Benzyl  com- 
pound re- 
covered as 
benzoic  acid 

hrs. 

mg. 

mg. 

mg. 

mg. 

mg. 

fo 

I 

24 

— 

95.2 

5.9 

89.3  " 

-- 

-- 

II 

8 

Benzyl 
sue . 
1500.0 

156. 

3.9 

152.1 

1228.2 

4.9 

III. 

16 

— 

497.8 

5.9 

491.9 

-- 

32.8 

IV. 

24 

-- 

417.3 

5.9 

411.4 

-- 

25.75 

V. 

24 

-- 

175.7 

5.9 

169.8 

— 

6.6 

VI. 

8 

Sodium 

benzoate 

1449.7 

1024.8 

9.9 

1014.9 

1228.2 

75.7 

VII. 

16 

-- 

205.0 

11.7 

193.3 

-- 

8.94 

VIII. 

24 

-- 

105.4 

11.7 

93.7 

— 

.83 

IX. 

8 

Benzyl 

alcohol 

1087.3 

863.8 

5.9 

857.4 

1228.2 

62.60 

X. 

16 

— 

285.6 

3.9 

281.7 

-- 

15.50  : 

XI. 

24 

— 

117.1 

5.9 

111.2 

— 

1.80 

XII. 

8 

Benzyl 

acetate 

1501.1 

863.8 

5.9 

857.9 

1228.2 

62  • 60 

XIII. 

16 

-- 

448.5 

-- 

-- 

-- 

28.77 

XIV. 

24 

214.8 

9.74 

Rabbit 

Daily 

9 

diet : 

male 

150  cc.  milk 

Table  XI. 
and  10  g.  sucrose 

weight:  2. 

and  15  g.  oats 

19. 

61  kg. 

Per- 

iod 

Dura- 

tion 

Benzyl  de- 
rivative 
used 

Benzoic  Acid 
To;bal 

Benzyl  de- 
rivative 
used  cal- 
culated as 
benzoic  ac. 

Benzyl 
compound 
recovered 
as  benzoic 
acid 

hr  s. 

mg. 

mg. 

mg. 

I. 

24 

— 

109.1 

— 

— 

II. 

8 

Benzyl  suc- 
cinate 
2000 

923.1 

1637.6 

49.7 

III. 

16 

-- 

338.9 

— 

14.04 

IV. 

24 

-- 

194.7 

-- 

5.25 

V. 

24 

-- 

-- 

-- 

-- 

VI. 

8 

Sodium 

benzoate 

1932.9 

763.5 

1637.6 

39.9 

VII. 

16 

-- 

219.6 

-- 

6.76 

VIII. 

24 

-- 

131.3 

-- 

1.35 

IX. 

24 

— 

-- 

-- 

— 

V 

• 

8 

Benzyl 

alcohol 

1449.7 

728.4 

1637.6 

37.8 

XI. 

16 

543.4 

— 

26.57 

XII. 

24 

— 

346.9 

-- 

14.52 

XIII. 

24 

-- 

— 

-- 

— 

XIV. 

* 

8 

* 

Benzyl 

acetate 

2012.3 

339.2 

1637.6 

14.07 

* Died 

• 

V 


20. 

Table  XII. 

Rabbit 

10 

Female 

v/ei  ght : 

1 • 5 • 

Daily 

diet : 150  cc . milk 

and  10  g.  sucrose 

and  10  g. 

oats . 

Period 

Dura- 

tion 

Benzyl  de- 
rivative 
used 

Benzoic  acid 

Benzyl  de- 
rivative 
used  cal- 
culated as 
benzoic  a. 

Benzyl  com- 
pound recov- 
ered as  ben- 
zoic acid. 

hrs. 

mg. 

mg. 

mg. 

/o 

I. 

24 

-- 

139.1 

-- 

-- 

II. 

8 

Benzyl 
sue . 
1000 

454.8 

818.8 

38.6 

III. 

16 

-- 

358.7 

-- 

26.8 

IV. 

24 

-- 

138 . 7 

— 

-- 

V. 

24 

-- 

-- 

-- 

— 

VI. 

8 

Sodium 

benzoate 

966.5 

616.6 

818.8 

58.4 

VII. 

16 

-- 

116.7 

-- 

-- 

VIII. 

24 

-- 

65.5 

-- 

-- 

IX. 

8 

Benzyl 

alcohol 

724.9 

354.6 

818.8 

35.3 

X. 

16 

-- 

416.2 

— 

42.19 

XI. 

24 

— 

65.5 

-- 

-- 

XII. 

8 

Benzyl 

acetate 

1006.7 

381.5 

818.8 

38.7 

XIII. 

16 

-- 

185.0 

-- 

14.6 

XIV. 

24 

119.5 

6.6 

. 


. 


* 


21. 


Tables  XIII 

Comparison  of  the  elimination  of  benzyl  succinate,  benzyl 
alcohol,  benzyl  acetate  and  sodium  benzoate. 


Rate 

of 

el ini nation 

Completeness 

of 

elimination 

Rabbit 

no . 

1 

2 

3 

4 

8 

o 

mt 

10 

1 

2 

3 

4 

8 

9 

10 

Benzyl 

cinate 

sue- 

3 

2 

2 

5 

4 

1 

3 

1 

1 

1 

3 

4 

2 

2 

Benzyl 

cohol 

al- 

1 

4 

1 

o 

3 

4 

3 

4 

1 

2 

1 

1 

Benzyl 

tate 

ace- 

2 

1 

3 

2 

3 

4 

2 

2 

2 

3 

2 

1 

4 

3 

Sodium 

zoate 

ben- 

1 

1 

2 

1 

2 

3 

3 

4 

0 " & 1 


. 


. 


22 


VII. 

Bibliography. 

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(2)  Macht  D.I.--J.  of  Pharm.  and  Exp.  Eher.  1918  XI,  263-7. 

(3)  Macht  D.I.  and  llelson  D.X. — Proc.  3oc.  Exp.  Biol.  led.,  1918, 

XVI.  25-6 

(4)  Macht  D.I.  and  Satani  Y.--J.  Pharm.  Exp.  Eher . Proc.,  1920, 

XV. , 244 

(5)  Macht  D.I.,  Satani  Y.  and  Schwartz  E.0.--J.  Urol.,  1921,  IV. 

355-61 

(6)  Macht  D.I.--J.  Pharm.  and  Exp.  Iher.  1919,  XIII,  509 

(7)  Sollnan  ?. — J.  Pharm.  a. id  Exp.  Ther . 1919  XIII  355-60 

(8)  Voegtlin  C.  and  Livingstone  A.E.--J.  Pharm.  and  Exp.  Eher. 

Proc.  1919,  XIII  515-4 

(9)  Mason  E.C.  and  Pieck  C.E.--J.  Lab.  and  Clin.  Med.,  1920, 

VI.  62-77 

(10)  Heller  E.A.  and  Steinfield  £. — H.Y.Med.J.,  1920,  CXI I,  160-1 

(11)  Hielson  C.  and  Higgins— J. A.— J.  Lab.  and  Clin.  Med.,  1921, 

VI.  388-92 

(12)  Hielson  C.  and  Higgins  J.A.--J.  Lab.  and  Clin.  .led.,  19-.1, 

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(13)  Macht  D.I. --Proc.  Soc.  Exp.  3iol.  Med.  1921,  XVIII,  177-9 

(14)  Volwiler  E.H.  and  Vliet  E.B. --J.A.  C . 3.  1921,  .^IIT  , 16  1 

(15)  Shonle  H.A.  and  Row  P.G.  — J.A.C.S.  1921,  XLIII,  361-5 

( 16 ) Kingsbury  F.B.  and  Swanson  — J.B.C.  1921,  .al .III,  1 - a1 

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